苦参碱对角叉菜胶诱导的关节炎大鼠炎症反应及血小板中精氨酸酶和腺苷核苷酸水解酶活性的影响

目的:研究苦参碱对角叉菜胶诱导的关节炎大鼠炎症反应及血小板中精氨酸酶和腺苷核苷酸水解酶活性的影响。方法:将大鼠随机分为正常组、模型组、阳性药物对照组(0.2 mg·kg^-1地塞米松)、苦参碱(低、中、高剂量)(20 mg·kg^-1、40 mg·kg^-1、80 mg·kg^-1)组。采用连续7 d关节内注射20μL的1%角叉菜胶构建类风湿关节炎大鼠模型,从第8天起,每天灌胃相应的药物,连续21 d。在给药第0、7、14、21天分别用游标卡尺测量各大鼠爪厚度,并进行关节炎指数评分;HE染色观察软骨损伤;流式细胞术测定大鼠外周血T细胞的功能表型;ELISA法测定大鼠血清中干扰素-γ(interferon-gamma,IFN-γ)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-4(interleukin-4,IL-4)和白细胞介素-10(interleukin-10,IL-10)的水平;通过Ehrlich试剂反应产生的尿素浓度来评估精氨酸酶活性;通过硫代巴比妥酸反应检测脂质过氧化;测定NTPD酶、5′-核苷酸酶活性和腺苷脱氨酶活性;蛋白印迹法测T细胞和软骨组织p65和p-IKBα/IKBα蛋白相对表达。结果:苦参碱高剂量组降低角叉菜胶诱导的关节炎大鼠爪厚度和关节炎指数,改善软骨损伤,降低Th1细胞与Th2细胞比例,降低IFN-γ+、IL-1β、IL-8、TNF-α含量,升高IL-4和IL-10含量,降低精氨酸酶活性、TBARS生成及ATP/ADP/AMP水解,增加腺苷脱氨酶活性,降低p65和p-IKBα/IKBα表达。结论:苦参碱能明显改善小鼠关节炎,这可能与其抑制炎症、维持Th1/Th2平衡、降低精氨酸酶和腺苷核苷酸水解酶活性、抑制NF-κB信号传导有关。

Effect of Matrine on Inflammatory Response and Activity of Arginase and Adenosine Nucleotide Hydrolase in Plateletsin Rats with Carrageenan-induced Arthritis

Objective:To study the effects of matrine on the inflammatory response and the activities of arginase and adenosine nucleotide hydrolase in platelets of carrageenan-induced arthritis rats.Methods:The rats were randomly divided into normal group,model group,positive drug control group(0.2 mg·kg^-1 dexamethasone),matrine(low,medium and high dose)(20 mg·kg^-1,40 mg·kg^-1,80 mg·kg^-1)group.A rat model of rheumatoid arthritis was constructed by intra-articular injection of 20μL 1%carrageenan for 7 consecutive days.From the 8 th day,the corresponding drugs were intragastrically administered every day for 21 consecutive days.On the 0 th,7 th,14 th,and 21 st days of administration,the thickness of each rat′s paw was measured with vernier calipers,and the arthritis index was scored;the cartilage damage was observed by HE staining;the functional phenotype of rat peripheral blood T cells was determined by flow cytometry;ELISA method to determine rat serum interferon-gamma(IFN-γ),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-4(IL-4)and interleukin-10(IL-10)levels;arginase activity is assessed by the concentration of urea produced by the Ehrlich reagent reaction;detection of lipid peroxidation by thiobarbituric acid reaction,determination of NTPD enzyme,5′-nucleotidase activity and adenosine deaminase activity;the protein expression of p65 and p-IKBα/IKBαin T cells and cartilage tissue was measured by western blot.Results:The high-dose matrine group reduced the paw thickness and arthritis index of carrageenan-induced arthritis rats,improved cartilage damage,reduced the ratio of Th1 cells to Th2 cells,and reduced IFN-γ+,IL-1β,IL-8 and TNF-αcontent,increase IL-4 and IL-10 content,reduce arginase activity,TBARS production and ATP/ADP/AMP hydrolysis,increase adenosine deaminase activity,reduce P65 and p-IKB/IKB expression.Conclusion:Matrine can inhibit arthritis,which may be related to its ability to inhibit inflammation,maintain Th1/Th2 balance,reduce arginase and adenosine nucleotide hydrolase activities,and inhibit NF-κB signal transduction.